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Outer membrane proteome of Actinobacillus pleuropneumoniae : LC‐MS/MS analyses validate in silico predictions
Author(s) -
Chung Jacqueline W.,
NgThowHing Christopher,
Budman Lorne I.,
Gibbs Bernard F.,
Nash John H. E.,
Jacques Mario,
Coulton James W.
Publication year - 2007
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200600979
Subject(s) - proteome , actinobacillus pleuropneumoniae , in silico , bacterial outer membrane , biology , microbiology and biotechnology , proteomics , computational biology , genome , actinobacillus , membrane protein , escherichia coli , bacteria , biochemistry , gene , serotype , genetics , membrane
The Gram‐negative bacterial pathogen Actinobacillus pleuropneumoniae causes porcine pneumonia, a highly infectious respiratory disease that contributes to major economic losses in the swine industry. Outer membrane (OM) proteins play key roles in infection and may be targets for drug and vaccine research. Exploiting the genome sequence of A. pleuropneumoniae serotype 5b, we scanned in silico for proteins predicted to be localized at the cell surface. Five genome scanning programs (Proteome Analyst, PSORT‐b, BOMP, Lipo, and LipoP) were run to construct a consensus prediction list of 93 OM proteins in A. pleuropneumoniae . An inventory of predicted OM proteins was complemented by proteomic analyses utilizing gel‐ and solution‐based methods, both coupled to LC‐MS/MS. Different protocols were explored to enrich for OM proteins; the most rewarding required sucrose gradient centrifugation followed by membrane washes with sodium bromide and sodium carbonate. This protocol facilitated our identification of 47 OM proteins that represent 50% of the predicted OM proteome, most of which have not been characterized. Our study establishes the first OM proteome of A. pleuropneumoniae .